HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Supplemental Figures All Versions of this Article: 2003-0106v1 24/4/896    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints/Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Van Damme, A. Articles by Chuah, M. K.L. Search for Related Content PubMed PubMed Citation Articles by Van Damme, A. Articles by Chuah, M. K.L.

TECHNOLOGY DEVELOPMENT

Efficient Lentiviral Transduction and Improved Engraftment of Human Bone Marrow Mesenchymal Cells

^a Center for Transgene Technology and Gene Therapy, Flanders Interuniversity Institute for Biotechnology and
^b Laboratory for Skeletal Development and Joint Disorders, University of Leuven, Leuven, Belgium;
^c Queen’s University, Kingston, Ontario, Canada;
^d Department of Pathology, Brown Medical School/Miriam Hospital, Providence, Rhode Island, USA

Key Words. Mesenchymal cell • Lentiviral • Bone marrow

Correspondence: Marinee K.L. Chuah, Ph.D. or Thierry VandenDriessche, Ph.D., Center for Transgene Technology and Gene Therapy, Flanders Interuniversity Institute for Biotechnology, University of Leuven, Herestraat 49, Building O&N1, Leuven B-3000, Belgium. Telephone: 32-16-346144; Fax: 32-16-345990; e-mail: marinee.chuah{at}med.kuleuven.be or thierry.vandendriessche{at}med.kuleuven.be

Received August 12, 2003; accepted for publication December 5, 2005.
Human bone marrow (BM) mesenchymal stem/progenitor cells are potentially attractive targets for ex vivo gene therapy. The potential of lentiviral vectors for transducing BM mesenchymal cells was examined using a self-inactivating vector that expressed the green fluorescent protein (GFP) from an internal cytomegalovirus (CMV) promoter. This vector was compared with oncoretroviral vectors expressing GFP from the CMV promoter or a modified long-terminal repeat that had been optimized for long-term expression in stem cells. The percentage of GFP-positive cells was consistently higher following lentiviral versus oncoretroviral transduction, consistent with increased GFP mRNA levels and increased gene transfer efficiency measured by polymerase chain reaction and Southern blot analysis. In vitro GFP and FVIII expression lasted for several months post-transduction, although expression slowly declined. The transduced cells retained their stem/progenitor cell properties since they were still capable of differentiating along adipogenic and osteogenic lineages in vitro while maintaining high GFP and FVIII expression levels. Implantation of lentivirally transduced human BM mesenchymal cells using collagen scaffolds into immunodeficient mice resulted in efficient engraftment of gene-engineered cells and long-term transgene expression in vivo. These biocompatible BM mesenchymal implants represent a reversible, safe, and versatile protein delivery approach because they can be retrieved in the event of an unexpected adverse reaction or when expression of the protein of interest is no longer required. In conclusion, efficient gene delivery with lentiviral vectors in conjunction with the use of bioengineered reversible scaffolds improves the therapeutic prospects of this novel approach for gene therapy, protein delivery, or tissue engineering.

This article has been cited by other articles:

				H. Matsui, M. Shibata, B. Brown, A. Labelle, C. Hegadorn, C. Andrews, R. P. Hebbel, J. Galipeau, C. Hough, and D. Lillicrap Ex Vivo Gene Therapy for Hemophilia A That Enhances Safe Delivery and Sustained In Vivo Factor VIII Expression from Lentivirally Engineered Endothelial Progenitors Stem Cells, October 1, 2007; 25(10): 2660 - 2669. [Abstract] [Full Text] [PDF]

				P. Koch, H. Siemen, A. Biegler, J. Itskovitz-Eldor, and O. Brustle Transduction of human embryonic stem cells by ecotropic retroviral vectors Nucleic Acids Res., October 6, 2006; 34(18): e120 - e120. [Abstract] [Full Text] [PDF]

				S. F. De Meyer, K. Vanhoorelbeke, M. K. Chuah, I. Pareyn, V. Gillijns, R. P. Hebbel, D. Collen, H. Deckmyn, and T. VandenDriessche Phenotypic correction of von Willebrand disease type 3 blood-derived endothelial cells with lentiviral vectors expressing von Willebrand factor Blood, June 15, 2006; 107(12): 4728 - 4736. [Abstract] [Full Text] [PDF]